Label-Free Multiphoton Microscopy: The Origin of Fluorophores and Capabilities for Analyzing Biochemical Processes

• Published in: Biochemistry (Moscow) Vol. 84; no. Suppl 1; pp. 69 - 88
• Main Authors: Shirshin, E. A., Yakimov, B. P., Darvin, M. E., Omelyanenko, N. P., Rodionov, S. A., Gurfinkel, Y. I., Lademann, J., Fadeev, V. V., Priezzhev, A. V.
• Format: Journal Article
• Language: English
• Published: Moscow Pleiades Publishing 2019; Springer; Springer Nature B.V
• Subjects: Analysis; Biochemical Phenomena; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Cells - metabolism; Chemical compounds; Connective Tissue - metabolism; Connective tissues; Diagnostic imaging; Equipment and supplies; Fluorescence; Fluorescent Dyes - chemistry; Fluorophores; Humans; Image processing; Imaging; Labels; Life Sciences; Methods; Microbiology; Microscope and microscopy; Microscopy; Microscopy, Fluorescence, Multiphoton - methods; Optical communication; Optical Imaging - methods; Review; Spatial discrimination; Spatial resolution; Russia; fluorophores; optical harmonic generation; FLIM; autofluorescence; molecular imaging; multiphoton microscopy
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1134/S0006297919140050

Multiphoton microscopy (MPM) is a method of molecular imaging and specifically of intravital imaging that is characterized by high spatial resolution in combination with a greater depth of penetration into the tissue. MPM is a multimodal method based on detection of nonlinear optical signals–multiphoton fluorescence and optical harmonics–and also allows imaging with the use of the parameters of fluorescence decay kinetics. This review describes and discusses photophysical processes within major reporter molecules used in MPM with endogenous contrasts and summarizes several modern experiments that illustrate the capabilities of label-free MPM for molecular imaging of biochemical processes in connective tissue and cells.